IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Hemisynthesis, NMR Characterization and UHPLC-Q-Orbitrap /MS² identification of (+)-Catechin oxidation products in red wines and grape seed extracts

Hemisynthesis, NMR Characterization and UHPLC-Q-Orbitrap /MS² identification of (+)-Catechin oxidation products in red wines and grape seed extracts

Abstract

(+)-Catechin—laccase oxidation dimeric standards were hemi-synthesized using laccase from Trametes versicolor in a water-ethanol solution at pH 3.6. Eight fractions corresponding to eight potential oxidation dimeric products were detected. The fractions profiles were compared with profiles obtained with two other oxidoreductases: polyphenoloxidase extracted from grapes and laccase from Botrytis cinerea. The profiles were very similar, although some minor differences suggested possible dissimilarities in the reactivity of these enzymes. Five fractions were then isolated and analyzed by 1D and 2D NMR spectroscopy. The addition of traces of cadmium nitrate in the samples solubilized in acetone-d6 led to fully resolved NMR signals of phenolic protons, allowing the unambiguous structural determination of six reaction products, one of the fractions containing two enantiomers. These products were then analyzed in grape seed extracts and red wines (UHPLC-Q-Orbitrap MS). The different dimers had different fragmentation patterns according to their interflavan linkage position. Oxidation dimeric compounds had a specific fragment ion at m/z 393, missing for B-Type dimers fragmentations. A fragment ion at m/z 291 occurred and was specific for oxidation dimeric compounds with a C-O-C linkage. Higher level oxidation products had abundant specific fragments: m/z 425, 397 and 245. These fragmentations were useful to identify them in complex samples such as grape seed extracts and wines. Three grape varieties and three ripening stages were selected and the corresponding seed extracts were obtained. The analyses revealed an increasing trend for the oxidation markers during grape ripening. The analysis of Syrah wines (2018, 2014, 2010) showed a decreasing trend of these molecules during wine ageing which might be due to further oxidation.

DOI:

Publication date: June 23, 2022

Issue: IVAS 2022

Type: Article

Authors

Saucier Cedric1, Deshaies Stacy1, Le Guernevé Christine1,2, Sommerer Nicolas1,2, Garcia Lucas Suc François1, Mouls Laetitia1

1SPO, Université de Montpellier, INRAE, Institut Agro, UMR SPO, Faculté de Pharmacie, 15 avenue Charles Flahault, 34000 Montpellier, France
2INRAE, PROBE Research Infrastructure, PFP Polyphenol Facility, 34060 Montpellier, France

Contact the author

Keywords

wine, grape, polyphenol,oxidation, catechin

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

The effects of alternative herbicide free cover cropping systems on soil health, vine performance, berry quality and vineyard biodiversity in a climate change scenario in Switzerland

There is an urgent need in viticulture to adopt alternative herbicide-free soil management strategies to mitigate climate change, increase biodiversity, reduce plant protection products and improve soil quality while minimizing detrimental effects on grapevine’s stress tolerance and fruit quality. To propose sustainable solutions, adapted to different pedoclimatic conditions in Switzerland, we developed a multidisciplinary 4-year project, started in 2020. Objectives of the project are to a) evaluate the impact of green covers (spontaneous flora, winter cover crop and permanent ground cover) on environmental and agronomic parameters and b) develop subsequently innovative strategies for different viticultural contexts of Switzerland. The project is divided into 3 phases: 1) diagnosis, 2) on-farm and 3) on-station experiments. Phase 1) consisted in an assessment of 30 commercial vineyards all over Switzerland, where growers already use different herbicide-free soil management strategies. The most promising practices identified in this exploratory phase will be replicated in commercial vineyards across Switzerland (“on-farm”) as well as in a classical randomized block design in an experimental plot (“on-station”). For phase 1), measurements consisted in evaluation of soil status (compaction, structure, roots development), soil microbial diversity (metagenomics), plant diversity and biomass, vine physiology (water stress, vigor, leaf nitrogen) and berry quality (acidity, sugar, available nitrogen). Interestingly, the permanent ground cover resulted in a higher Shannon index thus a higher biodiversity as compared to the other itineraries. The winter cover crop increased vine nitrogen and vigor while deteriorating soil quality, leaving the soil more exposed and compacted likely due to more frequent tillage. The spontaneous flora led to higher berry sugar accumulation, less nitrogen and higher malic acid concentration putatively due to a higher water retention of the flora in a particularly wet vintage. Phases 2) and 3) are required to confirm those tendencies, over the 3 next vintages and different climatic conditions.

Leaf vine content in nutrients and trace elements in La Mancha (Spain) soils: influence of the rootstock

The use of rootstock of American origin has been the classic method of fighting against Phylloxera for more than 100 years. For this reason, it is interesting to establish if different rootstock modifies nutrient composition as well as trace elements content that could be important for determining the traceability of the vine products. A survey of four classic rootstocks (110-Richter, SO4, FERCAL and 1103-Paulsen) and four new ones (M1, M2, M3 and M4) provided by Agromillora Iberia. S.L.U., all of them grafted with the Tempranillo variety, has been carried out during 2019. The eight rootstocks were planted in pots of 500 cc, on three soils with very different characteristics from Castilla-La Mancha (Spain). In the month of July, the leaves were collected and dried in a forced air oven for seven days at 40ºC. Then, the samples were prepared for the analysis determination, carried out by X-Ray fluorescence spectrometry. The results obtained showed that in the case of content in mineral elements in leaf, separated by soil type, we can report the importance of few elements such as Si, Fe, Pb and, especially, Sr. The rootstock does not influence the composition of the vine leaf for the studied elements that are the most important in determining the geochemical footprint of the soil. The influence of the soil can be discriminated according to some elements such as Fe, Pb, Si and, especially, Sr.

Influence of grapevine rootstock/scion combination on rhizosphere and root endophytic microbiomes

Soil is a reservoir of microorganisms playing important roles in biogeochemical cycles and interacting with plants whether in the rhizosphere or in the root endosphere. The composition of the microbial communities thus impacts the plant health. Rhizodeposits (such as sugar, organic and amino acids, secondary metabolites, dead root cells …) are released by the roots and influence the communities of rhizospheric microorganisms, acting as signaling compounds or carbon sources for microbes. The composition of root exudates varies depending on several factors including genotypes. As most of the cultivated grapevines worldwide are grafted plants, the aim of this study was to explore the influence of rootstock and scion genotypes on the microbial communities of the rhizosphere and the root endosphere. The work was conducted in the GreffAdapt plot (55 rootstocks x 5 scions), in which the 275 combinations have been planted into 3 blocks designed according to the soil resistivity. Samples of roots and rhizosphere of 10 scion x rootstock combinations were first collected in May among the blocks 2 and 3. The quantities of bacteria, fungi and archaea have been assessed in the rhizosphere by quantitative PCR, and by cultivable methods for bacteria and fungi. The communities of bacteria, fungi and arbuscular mycorrhizal fungi (AMF) was analyzed by Illumina sequencing of 16S rRNA gene, ITS and 28S rRNA gene, respectively. The level of mycorrhization was also evaluated using black ink coloration of newly formed roots harvested in October. The level of bacteria, fungi and archaea was dependent on rootstock and scion genotypes. A block effect was observed, suggesting that the soil characteristics strongly influenced the microorganisms from the rhizosphere and root endosphere. High-throughput sequencing of the different target genes showed different communities of bacteria, fungi and AMF associated with the scion x rootstock combinations. Finally, all the combinations were naturally mycorrhized. The root mycorrhization intensity was influenced by the rootstock genotype, but not by the scion one. Altogether, these results suggest that both rootstock and scion genotypes influence the rhizosphere and root endophytic microbiomes. It would be interesting to analyze the biochemical composition of the rhizodeposition of these genotypes for a better understanding of the processes involved in the modulation of these microbiomes. Moreover, crossing our data with the plant agronomic characteristics could provide insights into their roles on plant fitness.

Combining effect of leaf removal and natural shading on grape ripening under two irrigation strategies in Manto negro (Vitis vinifera L.)

The increasingly frequent heat waves during grape ripening pose challenges for high quality wine grape production. Defoliation is a common practice that can improve the control of diseases in bunches, but also it increases the exposure to sunlight. Grapes exposed to solar radiation reach temperatures over the optimum for berry development and maturation. This makes the development of irrigation and canopy management techniques of great importance to maximize yield and grape quality. A field experiment was carried out during 2021 using Manto negro wine grapes to study the effect of applied irrigation and different light exposure levels on grape quality. Two irrigation treatments were imposed based on the frequency and amount of water doses in a four-block experimental vineyard at Bodega Ribas (Mallorca). Three light exposure treatments were randomly applied in each irrigation plot. The light treatments included exposed clusters from pea size, non-exposed clusters, and shaded clusters after softening. Leaf area index and canopy porosity was estimated every 2 weeks. Midday leaf water potential was measured weekly. Additionally, apparent electrical conductivity was measured between rows to estimate the soil water content variability. Light and temperature sensors were installed at the bunch level to quantify the differences in bunch temperature and light intensity among treatments. The effect of irrigation and cluster light exposure on berry weight, TSS, TA, malic acid, tartaric acid, K+, and pH were analysed at 5 moments along grape ripening. During different heat waves, the natural shading technique decreased the maximum bunch temperature around 10 °C respect to the exposed bunches in both irrigation strategies. The combination of defoliation and shading techniques after softening decreased TSS at harvest and affected most of the quality parameters during the last stages of ripening, showing an interesting technique to delay ripening in warm viticulture areas.

Impact of climate variability and change on grape yield in Italy

Viticulture is entangled with weather and climate. Therefore, areas currently suitable for grape production can be challenged by climate change. Winegrowers in Italy already experiences the effect of climate change, especially in the form of warmer growing season, more frequent drought periods, and increased frequency of weather extremes.
The aim of this study is to investigate the impact of climate variability and change on grape yield in Italy to provide winegrowers the information needed to make their business more sustainable and resilient to climate change. We computed a specific range of bioclimatic indices, selected by the International Organisation of Vine and Wine (OIV), and correlated them to grape yield data. We have worked in collaboration with some wine consortiums in northern and central Italy, which provided grape yield data for our analysis.
Using climate variables from the E-OBS dataset we investigate how the bioclimatic indices changed in the past, and the impact of this change on grape productivity in the study areas. The climate impact on productivity is also investigated by using high-resolution convection-permitting models (CPMs – 2.2 horizontal resolution), with the purpose of estimating productivity in future emission scenarios. The CPMs are likely the best available option for this kind of impact studies since they allow a better representation of small-scale processes and features, explicitly resolve deep convection, and show an improved representation of extremes. In our study, we also compare CPMs with regional climate models (RCMs – 12 km horizontal resolution) to assess the added value of high-resolution models for impact studies. Further development of our study will lead to assessing the future suitability for vine cultivation and could lead to the construction of a statistical model for future projection of grape yield.